Substrate coated with a transparent organic film and manufacturing process

ABSTRACT

The present invention relates to a substrate coated with a transparent organic film, to a process for the manufacture of this substrate coated with the transparent organic film and to its use. The substrate coated with a film is characterized in that the film is an electrical insulator organic polymer which is transparent in at least one wavelength range and in that the said film is combined with a label which emits at least in the said wavelength range. It has an application in particular in a means for the detection of a chemical entity, for example a biochip, in a process for the quality control, a process for the certification or a process for the authentication of an object.

TECHNICAL FIELD

The present invention relates to a substrate coated with a transparentorganic film, to a process for the manufacture of this substrate coatedwith a transparent organic film, and to its use.

The term “transparent film” is understood to mean a film havingnoteworthy optical properties of transparency (absence of opticalabsorption, absence of “quenching”, and the like) over at least oneregion of the electromagnetic spectrum and in particular a low opticalextinction or “coefficient k” within the wavelength region underconsideration for the transparency. This optical extinction coefficientis measured, for example, by spectroscopic ellipsometry or usingspectrophotometry.

The substrate can be an insulating, conducting or semiconductingsubstrate with regard to electricity. It constitutes the support for thetransparent organic film and it is chosen according to the use or thedestination of the substrate coated with the transparent organic film inaccordance with the present invention.

The present invention has an application, for example, in the field ofthe quality control, of the certification or of the authentication ofsubstrates coated with thin transparent films.

This is because it makes it possible to prove the manufacture and/or theorigin of the substrate coated with the organic film of the presentinvention, in or on which it will have been possible to deliberatelyinsert a known fluorescent, phosphorescent or chemiluminescent label invery small amounts. In this application of the present invention, thesubstrate is any object.

It also has an application, for example, in the field of the detectionof the chemisorption or physisorption, on or in the transparent organicfilm, of chemical, biochemical or biological entities functionalizedbeforehand by a fluorescent, phosphorescent or chemiluminescent label,such as, for example, in processes for the detection by fluorescence ofchemical or biochemical analysis chips, such as DNA chips. In this typeof application, the substrate constitutes the support of the detectionmeans.

For example, in the case of a biochip, the substrate can, for example,be a support formed of silica, of gold or of a composite, such as Au/Si,Au/SiO₂ or more generally metal/substrate, and the transparent organicfilm can be one of the molecular means for attaching biological probesto certain parts of the surface.

When the biochip is brought into contact with a solution of sample to beanalyzed, pairings take place between the DNAs of the sample and thoseattached to the substrate. This attachment can, for example, be detectedby having labelled the DNAs of the sample beforehand with a fluorescent,phosphorescent or chemiluminescent label. In accordance with the presentinvention, the film is chosen in order to be transparent at the emissionwavelength of the fluorescent, phosphorescent or chemiluminescent labelused, so as to absorb as few as possible of the photons emitted by thislabel, to render it detectable at very low concentrations at the surfaceof the substrate and to minimize the interference with the measurement.The signal/noise ratio and the lower detection limits of the pairings onthe biochips are thus found to be improved thereby.

In the description which follows, the references in square bracketsrefer to the appended list of references.

PRIOR ART

As regards the biochip application, the documents FR-A-2 787 581 (1998),FR-A-2 787 582 (1998) and U.S. Pat. No. 5,810,989 (1998) disclose theelectrocopolymerization on a silica substrate of precursor monomers ofconducting polymers, such as pyrrole, with monomers functionalized byrecognition molecules, in particular oligonucleotides. This technique isamong the currently most widely used techniques for the localizedattachment of recognition molecules to the plots of a biochip.

In this technique, use is made of the adhesion of the conductingpolypyrrole film to the substrate, so as to carry out the attachmentthereto of the recognition molecules.

As disclosed in the abovementioned documents, and in the documentsFR-A-2 784 188 (1998) and FR-A-2 784 189 (1998), the chip thusfunctionalized is brought into contact with a solution of sample to beanalyzed comprising target molecules capable of coupling with therecognition molecules of the support.

In order to selectively detect the plots on which a coupling is presentbetween the recognition molecules and the target molecules, the lattercan advantageously be “labelled” with a fluorescent molecule, such as,for example, fluorescein or phycoerythrin, which exhibits an absorptionat 543 nm and an emission at 580 nm, the presence of which cansubsequently be detected using an appropriate optical device.

Unfortunately, polypyrrole has a not insignificant absorption in theemission wavelength region of the fluorescent label used. Thisdisadvantage is widely described in the literature relating to thetechniques concerned.

Specifically, Arwin et al., in the reference [1], measure an extinctioncoefficient k=0.3, for a refractive index n=1.45, on a polypyrrole filmwith a thickness of 22 nm on gold; Kim et al., in the reference [2],measure an extinction k=0.3 and an index n=1.6 at λ=632.8 nm, for apolypyrrole film with a thickness of 47 nm; Kim et al., in the reference[3], measure an index n=1.45 for an extinction k=0.28 on a polypyrrolefilm with a thickness of 54 nm in the oxidized state and an index n=1.6for an extinction k=0.21 on a film with a thickness of 47 nm in thereduced state; Guedon et al., in the reference [4], measure an indexn=1.7 for an extinction k=0.3 at 633 nm on a polypyrrole film on goldand for thicknesses of the film of between 7.5 and 20 nm.

Consequently, the polypyrrole used to attach the recognition moleculesabsorbs a large part of the fluorescence signal of the coupled targetmolecules. It thus interferes with the detection method.

In addition, as the target molecules and their labels are included inthe polypyrrole, in particular owing to the fact that the recognitionmolecules have been attached by copolymerisation, this interferingabsorption increases the value of the lower detection limit which can beachieved by this process.

In the field of diagnosis, for example, this disadvantage is a greatnuisance, since it is, specifically at low concentrations of targetmolecules, thus at low surface concentrations of fluorescent labels,that all the advantages of the biochip lie.

ACCOUNT OF THE INVENTION

An aim of the present invention is in particular to provide a substratecoated with a thin transparent film and a process for the manufacture ofthis substrate coated with the film which, first, responds to thetechnical problems posed in the prior art relating to biochips and,secondly, provides a novel powerful tool in the fields of the qualitycontrol, of the certification and of the authentication of any objects.

The substrate coated with a film of the present invention ischaracterized in that the film is an electrical insulator organicpolymer which is transparent in at least one wavelength range and inthat the said film is combined with a label which emits at least in thesaid wavelength range.

This is because the inventors have demonstrated that, unexpectedly, theelectrical conductor nature and the high optical extinction areconnected.

Thus, according to the present invention, the term “transparent” isintended to mean transparent at the detection wavelength of the labelused in accordance with the present invention.

According to a first embodiment of the present invention, the wavelengthrange in which the insulating polymer has to be transparent according tothe present invention is determined according to the label used, forexample, in the abovementioned applications.

In particular, once the label, and thus the emission wavelength of thelabel, has been chosen, it is easy to determine the value of theextinction of such and such a polymer at the desired wavelength, forexample by spectroscopic ellipsometry or using spectrophotometry, and toexamine whether it can be used according to the invention.

According to this first embodiment of the present invention, it is thusthe insulating polymer which is chosen as a function of the label.

According to a second embodiment of the present invention, after havingchosen an insulating polymer, the wavelength range within which it istransparent is determined and then, according to this wavelength range,a label which emits in the said transparency range of the polymer isselected.

According to this second embodiment of the present invention, it is thusthe label which is chosen as a function of the polymer.

According to the present invention, any insulating polymer is thereforecapable of being used as a transparent film over at least one wavelengthrange.

Mention may be made, among these, as nonlimiting examples, of vinylpolymers, vinyl copolymers and their blends, which may or may not becrosslinked, and in particular of polymers, copolymers and their blends,which may or may not be crosslinked, of acrylonitrile, ofmethacrylonitrile, of methyl methacrylate, of ethyl methacrylate, ofpropyl methacrylate, of butyl methacrylate, of hydroxyethylmethacrylate, of hydroxypropyl methacrylate, of cyano acrylate, ofacrylic acid, of methacrylic acid, of styrene, of para-chlorostyrene, ofN-vinylpyrrolidone, of vinyl halides, of acryloyl chloride or ofmethacryloyl chloride.

Mention may also be made, as nonlimiting examples, of the crosslinked ornon-crosslinked polymers chosen from polyacrylamides, polymers ofisoprene, of ethylene, of propylene, of ethylene oxide and moleculescomprising strained rings, of lactic acid or of its oligomers, oflactones, of ε-caprolactone or of glycolic acid, aspartic acid,polyamides, polyurethanes, parylene and polymers based on substitutedparylene, oligopeptides and proteins, and the prepolymers, macromers ortelechelics based on these polymers, and the copolymers and/or blendswhich can be formed from the monomers of these polymers or from thesepolymers themselves.

The choice of the insulating polymer to be used for the applicationunder consideration, for example among the abovementioned insulatingpolymers, can subsequently be determined by considerations other thanthose strictly related to the optical properties of the material.

Thus, according to the invention, the polymer can be selected, withinthe range of the polymers which can be used according to the invention,for example from a polymer capable of adhering to a substrate, a polymercapable of being functionalized, a polymer having thermoelasticproperties, and the like.

Among the abovementioned insulating polymers, the inventors havedirected their attention more specifically at vinyl polymers becausethey can be easily manufactured by various types of reactions, forexample ionic or radical reactions, in particular as thin films, andbecause they can also be obtained by the electrochemical route and canbe electrografted to surfaces which are conducting or semiconductingwith regard to electricity.

The vinyl polymers are obtained by polymerization of monomers infollowing generic formula (I):

-   -   in which R₁, R₂, R₃ and R₄ are hydrogen atoms or organic groups        of any nature, for example from hydrocarbons chosen, for        example, from alkanes, alkenes or alkynes; for example amides,        aldehydes, ketones, carboxylic acids, esters, acid halides,        anhydrides, nitriles, amines, thiols, phosphates, ethers, homo-        or heterocyclic aromatics or any cyclic group comprising these        functional groups, and any group carrying several of these        functional groups;    -   or by polymerization of a mixture of different monomers        corresponding to the formula (I) above.

In the nonlimiting examples of the implementation of the presentinvention set out below, the optical properties of polymethacrylonitrile(PMAN), with R₁=R₂=H, R₃=CH₃ and R₄=CN, and of poly(methyl methacrylate)(PMMA), with R₁=R₂=H, R₃=CH₃ and R₄=C(═O)OCH₃, were examined. These twomonomers can result in insulating polymers electrografted to conductingsurfaces by electroreduction in an aprotic organic medium.

According to the invention, in a specific application, the vinylpolymers can be grafted, optionally in a selective and localized way, tothe conducting or semiconducting surfaces of a substrate byelectrografting vinyl monomers, which renders them advantageous assubstitutes for polypyrroles in a biochip application.

These polymers can be deposited on surfaces of any type following theapplication of the present invention by the various processes known tothe person skilled in the art for depositing thin polymer films, such asthe techniques of spin coating; of dipping; of vaporization underultra-high vacuum; of CVD; of surface chemical polymerization, asdisclosed, for example, in U.S. Pat. No. 4,421,569, U.S. Pat. No.5,043,226 or U.S. Pat. No. 5,785,791; of photochemical grafting ofpolymers to surfaces, as disclosed, for example, in Patent ApplicationsWO-A-9908717 and WO-A-9916907; of grafting of polymers under irradiationof particles or photons; of chemical grafting to a surface of oxide orof another polymer, either directly or via chemical coupling agents(such as thiols, silanes, and the like); of depositions following apolymerization brought about by initiators, in particular radicalinitiators, which depositions are obtained in situ by electrochemistry;and the like.

According to a specific embodiment of the invention, it is possible, forexample, to obtain a deposit of transparent polymer as a thin layer bypolarizing a conducting or semiconducting surface in a solution or in agel comprising in particular diazonium salts and monomers which can bepolymerized by the radical route.

According to an advantageous embodiment, such polymer films are graftedto the surface of the substrate in particular as thin films, that is tosay with thicknesses of less than one micrometer, for example of between1 and 100 nm.

Other thicknesses are possible in the context of the present inventionas defined in the appended claims.

The films can be either preformed polymer films grafted to the surface,for example by the chemical, electrochemical or photochemical route, inone or more stages, or films constructed directly on the surface fromprecursor monomers, for example initiated by the chemical,electrochemical or photochemical route.

This grafting can be carried out, according to the physicochemicalcharacteristics of the process employing the present invention, onsurfaces which are insulating, conducting or semiconducting with regardto electricity.

According to a favoured embodiment, ultrathin films of transparentpolymers can be obtained on surfaces which are conducting orsemiconducting with regard to electricity by electrografting vinylmonomers, for example as disclosed in Patent Application EP-A-038 244.

According to the present invention, the label can be any label providedthat it can be combined with an electrical insulator organic polymer inaccordance with the present invention, the said polymer having to betransparent in at least the emission wavelength range of the label.

For example, the label can be a fluorescent, phosphorescent orchemiluminescent label.

It can be, for example, a label chosen from fluorescein or substitutedfluoresceins, such as fluoresceindiacetate, 5- and6-carboxyfluoresceins, 5- and 6-carboxyfluoresceindiacetates, thesuccinimidyl ester of 5- and 6-carboxyfluoresceins, and the like;rhodamine and substituted rhodamines; coelenterazine and substitutedcoelenterazines; aequorin; luciferin and substituted luciferins;bromochloroindoxyl phosphates (BCIP); luminol; nonyl acridine orange(NAO); 5, 5′, 6, 6′-tetrachloro-1, 1′, 3,3′-tetraethylbenzimidazolyl-carbocyanine chloride;4-(4-ditetradecylaminostyryl)-N-methylpyridinium iodide;1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate;3,3′-dihexadecyloxacarbocyanine hydroxyethanesulfonate;bis-(1,3-dibarbituric acid)-trimethine oxanol; tetrazolium salts;calcium complexes; potassium complexes; anthraquinone; anthracene;pyrene; doxorubicin; phycoerythrin; porphyrins; phthalocyanines and moregenerally organometallic complexes; fluorescent proteins and inparticular GFPs, Green Fluorescent Proteins (K. F. Sullivan, S. A. Kay,“Methods in Cell Biology: Volume 58: Green Fluorescent Proteins”,Academic Press, 1999); salts of fluorescent minerals (and in particularuranium salts); and any molecule having a fluorophore group. Achemiluminescent label is a label which emits a fluorescence when it isbrought into the presence of another molecule. These labels are known toa person skilled in the art; it can be, for example, the biotin-avidinpair commonly used in molecular biology.

The amount of labels can be very low in view of the choice of thecombined electrical insulator organic polymer. For example, the labelcan be at a concentration of the order of the nanomolar to themicromolar, for example from 1 nM to 10 μM.

According to the invention, the term “combined label” is understood tomean a label mixed with the insulating organic polymer film, or attachedto the monomer(s), for example functionalized beforehand, used for themanufacture of the polymer film, or attached directly or indirectly tothe surface of the film, or trapped in the polymer film during themanufacture of the latter on the surface, or simply deposited on thefilm using a solution of the label.

For example, in the quality control, certification or authenticationapplications, the label can be mixed with the insulating organicpolymer, or inserted after deposition of the polymer by dipping in asolution of a solvent which swells the polymer comprising the label, orinserted during the synthesis by carrying out the polymerization in thepresence of the label in the synthesis medium, or inserted during thesynthesis by carrying out the polymerization with monomers or comonomerschemically functionalized with the label, or deposited on the polymerusing a solution of labels.

For example, in the applications relating to biochips, the label can becombined by directly grafting onto the film or by indirectly graftingonto recognition molecules grafted to the insulating organic polymerfilm.

Thus, the present invention makes it possible, for example, to solve theproblems of the abovementioned prior art relating to biochips bysubstituting the commonly used polypyrrole by an insulating polymer inaccordance with the present invention, for example by a vinyl polymer.This is because, as demonstrated below in the implementational examplesof the present invention, these insulating polymers have extinctionswhich are 10 to 100 times smaller than those of conducting polymers suchas polypyrrole, which greatly reduces their interference with a label indetection processes using a chip, for example a biochip.

Consequently, the present invention makes it possible to manufacturedetection chips which are much more sensitive than those of the priorart by lowering the lower detection limit, for example with respect tochips using polypyrrole or another conducting polymer.

These detection chips can be manufactured by any known means, exceptthat the conducting films deposited on the substrates will have to bereplaced by insulating films chosen in accordance with the presentinvention.

The present invention also relates to the use of such thin transparentorganic films.

Generally, such a coating is capable of accommodating a label, forexample a fluorescent, phosphorescent or chemiluminescent label, chosenwithin the region of the electromagnetic spectrum corresponding to itsarea of transparency and of allowing the detection of this label even,and in particular, when this label is present at very low concentrationsin or on the organic film, this being the situation by virtue of theoptical transparency properties of the insulating organic polymer filmchosen.

Thus, the present invention relates to the use of a film of electricalinsulator organic polymer which is transparent in at least onewavelength range combined with a label which emits at least in the saidwavelength range in a process for the detection of a chemical entity.

This is because the film of insulating organic polymer of the presentinvention can be functionalized with recognition molecules for thechemical entity, such as nucleic acids, proteins, antigens, antibodies,synthetic organic molecules, and the like. For example, in a process fordetection by biochip, the chemical entity can be DNA.

The film of insulating organic polymer of the present invention can alsobe used to encapsulate molecules, for example bioactive molecules, suchas doxorubicin, the molecular structure of which is such that thesemolecules are fluorescent. In this application, the molecule has twoproperties belonging to it alone: that related to its bioactivity andthat related to its fluorescent nature.

More generally, the present invention relates to the use of the film ofelectrical insulator organic polymer of the present invention which istransparent in at least one wavelength range combined with a label whichemits at least in the said wavelength range in a means for the detectionof a chemical entity.

As set out above, the detection means can be a biochip, such as a DNAchip, a protein chip, a chemical probe, and the like.

The present invention also relates to the use of the film of electricalinsulator organic polymer of the present invention which is transparentin at least one wavelength range combined with a label which emits inthe said wavelength range in a process chosen from a process for thequality control, a process for the certification or a process for theauthentication of an object.

In the quality control of an industrial process for the deposition ofthin insulating polymer films, for example, it is essential to be ableto characterize the thickness of the coating. On thin films, inparticular when these films have a thickness of less than 100 nm, it isnecessary to resort to expensive and slow techniques, such asprofilometry or ellipsometry, to obtain reliable measurements of thethicknesses. In addition, rapid measuring devices often have a lowerdetection limit of greater than one micron. Furthermore, when thesamples to be described are complex in shape (microbeads, meshes,powders, and the like), the measurements, for example by profilometry orellipsometry, are difficult.

By combining, according to the present invention, a polymer film and alabel, for example a fluorescent, phosphorescent or chemiluminescentlabel, it is possible to obtain, in a very simple way, an indirectmeasurement of the thickness of the film by measuring the intensity ofthe fluorescence emitted.

This can be carried out, for example, by subjecting the object on whichthe polymer film combined with the label according to the presentinvention has been deposited to irradiation by a light source, thespectrum of which contains at least the absorption wavelength of thefluorophore, and by measuring the intensity of the fluorescence emittedover the emission wavelength of the fluorophore.

For this, it is sufficient to prepare beforehand a calibration curve, orstandard curve, on which the intensity of fluorescence of a filmcombined with a fluorescent label is plotted as a function of thethickness of the film, measured by profilometry or ellipsometry, forvarious flat samples covered with the polymer film according to theindustrial deposition process in question, the label being combined withthe film at a chosen concentration identical for all samples.

From this standard curve, the measurement of the intensity offluorescence emitted by an identical polymer film manufactured by theindustrial process in question and comprising the chosen concentration,which is identical to that of the abovementioned samples, of label issufficient to determine whether the film exhibits the requiredthickness, that is to say whether it does or does not meet thespecifications in terms of thickness, from the knowledge of the area ofthe said object.

If this area is not known, the same protocol provides at least a meansfor the in-line monitoring of the reproducibility of the industrialmethod for the deposition of a polymer film.

The monitoring of the thickness of the films according to the presentinvention, must be carried out in a very short time from a standardcurve.

In this quality control application, the present invention makes use ofthe active properties of the coating because of the involvement of afluorophore.

The present invention can also be used in the certification or theauthentication of an object. For this, it is sufficient to deposit, onthe said object, a film of an electrical insulator organic polymer whichis transparent in at least one wavelength range combined with a labelemitting at least in the said wavelength range according to the presentinvention.

Thus, by simple measurement of fluorescence, it is possible to determinewhether the object is an authentic object, that is to say comprising thefilm combined with the label, or whether it is a copy of the saidobject.

The establishment of an infringement is often difficult, since numerousprocesses differ both in terms of the treated component and in thenature of the interface which has been constructed between the polymerfilm and the surface of the untreated component. As this interface isburied beneath the film which has been deposited, it becomes difficultto analyse it through the deposited film, in particular when itsthickness is greater than 10 nm. In this case, a proprietor of anintellectual property title relating to a process for the deposition ofthin films can advantageously label the films manufactured by him/herwith a label, for example a fluorescent, phosphorescent orchemiluminescent label, in accordance with the present invention in asufficiently low concentration for a measuring device such as thatdescribed above to be necessary for its detection.

The abovementioned applications are to be regarded only by way ofillustration and neither these applications nor the procedure by whichthe insulating polymers constituting the film are deposited on thesurface of the substrates should constitute a limitation to theapplication of the present invention.

This is because a person skilled in the art will know how to measure,with regard to other applications, the significance of the presentinvention by combining one or more organic coating(s) of one or moreelectrical insulator polymer(s) having low optical extinctions in awavelength range and a label emitting in the said range.

Other advantages and characteristics of the present invention willbecome more apparent to a person skilled in the art on reading theexamples below, given by way of illustration and without limitation,with reference to the appended figures.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 is a diagram illustrating the principle of the rotating polarizerellipsometry used to measure the thickness of the films of insulatingpolymer according to the present invention.

FIGS. 2 and 3 are ellipsometric measurement spectra of gold, assubstrate, for different angles of incidence in degrees: tan(Ψ)=f(λ(nm))(FIG. 2) and cos(Δ)=f(λ(nm)) (FIG. 3) (λ=wavelength in nm).

FIG. 4 is a graph combining measurements of index (I) and (N) and ofoptical extinction coefficient (E) and (K) carried out on a substrate ofgold alone (G) and on a substrate covered with an organic film, measuredat an angle of 75° for all the wavelengths between 300 and 800 nm.

FIGS. 5 and 6 are ellipsometric spectra of a platinum substrate withoutan organic polymer film obtained from measurements carried out over aspectral range extending from 300 to 800 nm with a step of 5 nm:tan(Ψ)=f(λ(nm)) with an incidence of 750 (FIG. 5) and cos(Δ)=f(λ(nm))with an incidence of 750 (FIG. 6).

FIG. 7 is a graph combining measurements of index (I) and (N) and ofoptical extinction coefficients (E) and (K) carried out on a platinumsubstrate without an organic film, measured at an angle of 75° for allthe wavelengths between 300 and 800 nm, and values given by the“Handbook of Optical Constants of Solids”, edited by E. D. Palik.

FIGS. 8 a), 9 a), 10 a and 11 a) represent ellipsometric spectra of aplatinum substrate covered with a PMAN film which are obtained frommeasurements carried out between 55 and 75° with a step of 5°, thespectral range extending from 300 to 800 nm with a step of 5 nm:tan(Ψ)=f(λ(nm)), for the samples AuMAN7, AuMAN24, Au2401 and Au2301respectively.

FIGS. 8 b), 9 b), 10 b) and 11 b) represent ellipsometric spectra of aplatinum substrate covered with a PMAN film obtained from measurementscarried out between 50 and 75° with a step of 5°, the spectral rangeextending from 300 to 800 nm with a step of 5 nm: with an incidence of75° (FIG. 5) and cos(Δ)=(λ(nm)), for the samples AuMAN7, AuMAN24, Au2401and Au2301 respectively.

FIG. 12 is a graphical representation of the measurements of thereflection (in %) of the samples with a gold substrate coated withdifferent transparent electrical insulator organic polymer filmsaccording to the invention as a function of the wavelength (in nm).

FIGS. 13 a) and 13 b) are spectra of ellipsometric measurements carriedout between 50 and 75° with a step of 5°, the spectral range extendingfrom 300 to 800 nm with a step of 5 nm, on a platinum substrate coatedwith a film of conducting organic polymer based on diazonium salts(sample 01010Pt6), for different angles of incidence in degrees:tan(Ψ)multiangles=f(λ(nm)) (FIG. 13 a)) and cos(Δ)multiangles=f(λ(nm))(FIG. 13 b)).

FIGS. 14 a) and b) are spectra of ellipsometric measurements carried outbetween 50 and 75° with a step of 5°, the spectral range extending from300 to 800 nm with a step of 5 nm, on a platinum substrate coated with afilm of conducting organic polymer based on diazonium salts (sample01010Pt14), for different angles of incidence in degrees:tan(Ψ)multiangles=f(λ(nm)) (FIG. 14 a)) and cos(Δ)multiangles=f(λ(nm))(FIG. 14 b)).

FIG. 15 represents the results of spectrophotometric measurementsintended to determine the reflection of a sample of platinum coated witha conducting organic film based on diazonium salts (samples 0101Pt6 and0101Pt14), over the range 400 to 800 nm with a reference wavelength of560 nm.

FIG. 16 represents the results of spectrophotometric measurementsintended to determine the losses of a sample of platinum coated with aconducting organic film based on diazonium salts (samples 0101Pt6 and0101Pt14), over the range 400 to 800 nm with a reference wavelength of560 nm.

FIG. 17 is a diagram for the dipolar modelling of a surface fluorophore:it represents a dipole with a dipole moment m placed above a surface x.Three media are distinguished: the environment of the dipole, the area 1with an index n₁, in this instance the superstrate; a stack of thinlayers, the area 2 with an index n₂; and the substrate, the area 3 withan index n₃.

FIGS. 18 a) and 18 b) represent modellings (signal (ua)=f(thickness ofthe polymer) (in nm)) of the fluorescence signal of a fluorophore dipoleplaced at the surface of an organic film according to two orientations,parallel (//) (FIG. 18 a)) and perpendicular (⊥) (FIG. 18 b)) to thesurface. In these figures, case 1: n=1.5 and k=0.02; case 2: n=1.5 andk=0.003; case 3: n=1.5 and k=0.0 (extreme value); the case 4 models atypical conducting organic film, such as those obtained from diazoniumsalts: n=1.5 and k=0.4. In these figures, the axis of the abcissaerepresents “t” the thickness of the polymer film and the axis of theordinates represents “s” the signal.

FIGS. 19 a) and 19 b) are diagrammatic representations of goldsubstrates coated, on the one hand, with a film of polypyrrole (FIG. 19a)) in accordance with the prior art and, on the other hand, with aninsulating organic polymer film (ManAu11) (FIG. 19 b)) in accordancewith the present invention.

FIG. 20 a) is a negative (objective ×50; exposure time: 200 ms) of thefluorescence of a drop (0.5 μl of a 1 μM solution) of Cydctp (trademark) fluorophore from Amersham deposited on a gold substrate, over thegold zone.

FIG. 20 b) is a negative (objective ×50; exposure time: 200 ms) of thefluorescence of a drop (0.5 μl of a 1 μM solution) of Cydctp (trademark) fluorophore from Amersham deposited on a film of polypyrroledeposited on a gold substrate, over the zone corresponding to thepolypyrrole.

FIG. 20 c) is a negative (objective ×50; exposure time: 200 ms) of thefluorescence of a drop (0.5 μl of a 1 μM solution) of Cydctp (trademark) fluorophore from Amersham deposited on a film ofpolymethacrylonitrile (MAN) deposited on a gold substrate (AuMan11sample), over the zone corresponding to the deposition of MAN.

EXAMPLES Example 1 Techniques for Measuring and Modelling the OpticalProperties of a Sample

The extinction measurements are carried out by spectroscopicellipsometry. This is a nondestructive optical technique which makes itpossible to characterize, by determination of the index and of thethickness, thin deposits of materials by making use of the modificationswhich the latter produce on the polarization of light.

When a beam is reflected at the surface of a sample, its state ofpolarization is modified. This is because, at oblique incidence, theelectrical field of a light wave is broken down along two specificdirections, one of which is perpendicular to the plane of incidence(wave S) and the other parallel to this plane (P). These two wavesinteract differently with the surface of a sample and reveal amplitudereflection coefficients r_(s) and r_(p) according to whether they relateto the S or P waves.

The change in state of polarization, which results from the differencein amplitude behaviour and phase behaviour of the S and P waves, canthen be characterized by ρ according to the following equation eq1:$\begin{matrix}{\rho = {\frac{r_{P}}{r_{S}} = {{\tan(\Psi)}{\mathbb{e}}^{{\mathbb{i}}\quad\Delta}}}} & {eq1}\end{matrix}$

-   -   tan(Ψ) represents an amplitude ratio and Δ a phase difference        between the S and P polarizations.

The ellipsometer used is the GESP5 (trade mark) from SOPRA. It makes itpossible to measure the parameters tan(Ψ) and cos(Δ) as a function ofthe wavelength, hence the term “spectroscopic ellipsometry”, and/or ofthe angle of incidence θ of the analytical light beam. The operatingprinciple of the rotating polarizer ellipsometer used is set out in FIG.1.

In this figure, S represents a light source, s and p the polarizationvectors of the incident light, P and A rotating polarizers and D adetector.

By means of modelling the sample, it is possible to calculate thecharacteristics of the latter using regression algorithms on themeasurements given by the ellipsometer.

The samples, thickness t, index n and extinction k, were characterizedby regression of the ellipsometric measurements using smoothing softwareby nonlinear regression in the complex plane with a nondispersive lawand then with a Lorentz oscillators model.

The consistency of the results was confirmed with the spectrophotometricmeasurements.

Example 2 Samples Examined

The measurements were carried out on eight different samples, thecharacteristics of which are summarized in Table 1.

Three separate series of samples were analysed:

-   -   a series of PMAN films electrografted to gold, with a thickness        varying between 9 and 150 nm. These films are obtained by        electroreduction of a 2.5M solution of methacrylonitrile in        anhydrous acetonitrile on a gold electrode in the presence of        0,05 M tetraethylammonium perchlorate (TEAP) as supporting        electrolyte. The electroreduction takes place under voltametric        conditions between −0.3 and −2.6 V/(Ag⁺/Ag) at 100 mV/s, in        nonseparated compartments, with a platinum counterelectrode with        a high surface area. The various thicknesses are obtained by        varying the number of voltametric sweeps;    -   two PMMA films electrografted to gold, one with a thickness of        100 nm, the other thick (thickness >0.5 μm). These films are        obtained by electroreduction of a methyl methacrylate solution        under the same conditions as for the PMAN films;    -   two films obtained by electroreduction of a 10⁻³M solution of        para-nitrophenyldiazonium (PNPD) tetrafluoroborate in anhydrous        acetonitrile on a platinum electrode, in the presence of 5×10−²M        TEAP as supporting electrolyte. The potential sweeps are applied        from +0.3 V/(Ag⁺/Ag) to −2.9 V/(Ag⁺/Ag), at −200 mV/s. Two films        with respective thicknesses of 3 and 30 nm are obtained by this        protocol.

This final series of samples was chosen as the organic films obtained byelectrografting PNPD are electrically conducting, in contrast to thefilms of the first two series. In addition, the ellipsometricmeasurements will also be compared to those obtained on a polypyrrolefilm as conducting polymer.

The thicknesses of the various films listed in Table I below aremeasured by profilometry. The arithmetic roughnesses, measured byprofilometry, according to two tip distances: 500 μm and 2 mm, are alsolisted in this table. TABLE I References and characteristics of thesamples used in the implementational examples Thick- Ra at Ra at Sub-ness 500 μm 2 mm Name strate Coating (nm) (nm) (nm) MANAu 15 GoldPolymethacrylonitrile 9 4   0.7 MANAu 24 Gold Polymethacrylonitrile 283.9 6.5 Au MAN 11 Gold Polymethacrylonitrile 50 2.9 2.9 Au MAN 7 GoldPolymethacrylonitrile 150 3.4 5.3 Au 2301 Gold Polymethacrylonitrile 100— — Au 2401 Gold Polymethylmethacrylate Thick — — 0101 Pt 14 PtNitrobenzoic 3 6.5 8.3 0101 Pt 6 Pt Nitrobenzoic 30 2.4 2.4

In this table, the polymethacrylonitrile or poly(methyl methacrylate)films are insulating and the nitrobenzoic films are conducting.

ELLIPSOMETRIC MEASUREMENTS ON THE SUBSTRATES

Characteristics of the Gold Layer (Substrate)

The ellipsometric spectra of the gold are measured for different anglesof incidence. In this way, it is possible to assess the quality of thefinal result as a function of the results obtained for these variousvalues, between 50° and 75° with a step of 5°. The measurement spectraextend between 300 and 800 nm with a measurement step of 5 nm. They arepresented in the appended FIGS. 2 and 3.

The measurement of the gold was carried out on the MANAu11 sample, onthe part not dipped in the reaction mixture and on which no organic filmhas been deposited.

The aim is to find the values of index and of extinction of this goldlayer; its thickness of greater than a micron makes it possible to putit in the same category (ellipsometric) as a substrate. An inversion,equation resolution, of the data is then carried out in order to extractthe n and k coefficients of the gold. This operation is carried out foreach measurement angle. The indices measured at an angle of incidence of75° have been listed in the appended FIG. 4 for all the wavelengthsbetween 300 and 800 nm.

For comparison, the measurements carried out on a substrate of goldalone have also been listed, independently of the samples carrying agrafted organic film, which makes it possible to monitor the stabilityof the measurements.

Characteristics of the Platinum Layer (Substrate)

The ellipsometric spectra of platinum are measured under the sameconditions as for gold: measurement angle between 50° and 75° with astep of 5° and the spectral range extends from 300 to 800 nm with a stepof 5 nm.

The measurement of the platinum was carried out on the 0101Pt6 sample,on the part not dipped in the reaction mixture and on which no organicfilm has been deposited.

FIGS. 5 ((tanΨ)=f(λ(nm)), incidence 75°) and 6 ((CosΔ)=f(λ(nm)),incidence 75°) are graphical representations of the results obtained forthese measurements.

The aim is to find the values of index and extinction of this platinumlayer: its thickness of greater than one micron allows it to be put intothe same category, from an ellipsometric viewpoint, as a substrate. Aninversion of the data of the measurement at 75° is then carried out inorder to extract therefrom the n (index) and the k (extinctioncoefficient). The results of index (I) and extinction (E) thus obtainedare combined in FIG. 7. Values given by the “Handbook of OpticalConstants of Solids”, edited by E. D. Palik, have also been representedin this graph.

For the characterizations of the samples, the results given by theellipsometric measurement will be taken as reference values forplatinum.

ELLIPSOMETRIC MEASUREMENTS OF THE SAMPLES

The measurements are carried out over the spectral range 300-800 nm andat θ=60°.

Ellipsometric Results of the Samples on a Gold Substrate

In order to be more exact with regard to the results, the AuMAN7,MANAu24, Au2301 and Au2401 samples were measured for variable angles.

The measurements carried out have made it possible to put together thespectra represented in FIGS. 8 a) and b) for AuMAN7; in FIGS. 9 a) andb) for AuMAN24; in FIGS. 10 a) and 10 b) for Au2401; and in FIGS. 11 a)and b) for Au2301. The a) figures correspond to the tan(Ψ)multianglemeasurements and the b) figures correspond to the cos(Δ)multianglemeasurements.

Spectrophotometric Results of the Samples on the Gold Substrate

The samples were measured with a manual “Lambda9m” (trade mark)spectrophotometer in order to determine the Reflection, the Transmissionand the Losses of each of them over the wavelength range 400-800 nm,with 560 nm as reference wavelength.

It is found that all the samples on gold have the same spectra and that,at 560 nm, 72.5%<R<84.2%, T=0 and 157%<L<27.5%. These results arecombined in Table 2 below: TABLE 2 Reflection (R), Transmission (T) andLosses (L) at 560 nm for the samples on gold at 560 nm Sample R(%) T(%)L(%) MANAu 11 72.55 0 27.46 MANAu 15 81.32 0.02 18.66 MANAu 24 75.450.03 24.51 AuMAN 7 79.96 0 20.05 Au 2301 77.54 0.0207 22.44 Au 240184.19 0.04 15.77

The appended FIG. 12 is a graphical representation of the measurementsof the reflection (in %) carried out on these samples comprising a goldsubstrate coated with different transparent and electrical insulatororganic polymer films according to the invention as a function of thewavelength (in nm).

It is found that all the samples on gold have the same spectra and that,at 560 nm, 72.5%<R<84.2%, T=0 and 15.7%<L<27.5%. It is found inparticular that these insulating films have few losses over a widewavelength range. This observation will become even clearer on comparingthe results obtained with the conducting PNPD films.

Ellipsometric Results of the Samples on a Platinum Substrate

FIGS. 13 a) and 13 b) represent the ellipsometric measurement spectrarecorded between 50 and 75° with a step of 5°, the spectral rangeextending from 300 to 800 nm with a step of 5 nm, on the 0101Pt6 samplesfor various angles of incidence in degrees: tan(Ψ)multiangles=f(λ(nm))(FIG. 13 a)) and Cos(Δ)multiangles=f(λ(nm)) (FIG. 13 b)).

FIGS. 14 a) and 14 b) represent the ellipsometric measurement spectrarecorded between 50 and 75° with a step of 5°, the spectral rangeextending from 300 to 800 nm with a step of 5 nm, on the 0101Pt14samples for various angles of incidence in degrees:tan(Ψ)multiangles=f(λ(nm)) (FIG. 14 a)) and Cos(Δ)multiangles=f(λ(nm))(FIG. 14 b)).

Spectrophotometric Results of the Samples on a Platinum Substrate

The samples were measured with a manual “Lambda9m” (trade mark)spectrophotometer in order to determine the Reflection, the Transmissionand the Losses of each of them over the wavelength range 400-800 nm,with 560 nm as reference wavelength.

It is found that the samples on platinum have very different spectra.The reflection, transmission and losses are summarized in Table 3 below.It is observed that the losses in the PNPD films, PNPD being conductingpolymers, are much more significant than in the case of insulating filmsproduced on gold substrates. TABLE 3 Reflection (R), Transmission (T)and Losses (L) at 560 nm for the samples on platinum at 560 nm SampleR(%) T(%) L(%) 0101Pt14 45.72 0.02 54.26 0101Pt6 57.32 0.02 42.66

FIG. 15 represents the reflection and FIG. 16 represents the losses ofthe platinum samples coated with a conducting polymer 0101Pt6 and0101Pt14 over the range 400 to 800 nm with a reference wavelength of 560nm.

CHARACTERIZATION OF THE SAMPLES

To characterize the samples (t, n and k), regression was carried out onthe ellipsometric measurements using SporX (trade mark) software with anondispersive law and then with Lorentz oscillators. Consistent resultswere obtained, on the one hand between the two models and, on the otherhand, with the spectrophotometric measurements.

The assessment of these characterizations is summarized in Table 4.

Very low extinction coefficients are measured for the insulating vinylfilms (k<0.02), whereas the conducting PNPD films give extinctions whichare at least 10 times higher.

By way of comparison, the polypyrrole films with a thickness similar tothose of the samples measured (20 nm) have an extinction coefficient of0.3 to 0.5, measured under the same conditions. TABLE 4 Opticalcharacteristics of the samples t t (profilo) (ellipso) Sample Nature nmnm n k MANAu15 insulating 9 5-6 1.5 0.02 MANAu24 insulating 28 24 1.5 0AuMAN11 insulating 50 40-46 1.5-1.6 0.003 AuMAN7 insulating 150 149 — —Au2301 insulating 100 — 1.1 0.02 Au2401 insulating (thick) — 1 0 0101Pt6conducting 3 — 1.9 0.1 0101Pt14 conducting 30 25-30 0.9 0.5

The aim now is to characterize the signal which would be emitted by afluorophore adsorbed on the surface of the film.

First of all, the expected advantage of the fact of the low extinctionsis presented and then it is shown, on a simple test, that thefluorescence yield is effectively better on the PMAN films on gold thanon a polypyrrole film of the same thickness.

The fluorophore is modelled as being an electric dipole, as representedin FIG. 17, that is to say the small source which can be envisaged inthe context of the electromagnetic theory. This step is not in any wayharmful to the general application of the question treated, since theGreen's function of the problem can be calculated and any more complexelectromagnetic source, such as superposition of base dipoles, can bereconstructed. Furthermore, it is assumed that the electromagneticfields will not be quantized. The comparison of theoretical models formodelling the fluorescence and the luminescence in laser cavities hasshown near-similar results, whether the viewpoint is classical orquantum.

On the basis of the results of Table 4, the following cases aresimulated: case 1: n=1.5/k=0.02; case 2: n=1.5/k=0.003; case 3:n=1.5/k=0 (extreme value); the final case models a typical conductingorganic film, such as, for example, the films obtained from diazoniumsalts (samples 0101Pt6 and 0101Pt14) case 4: n=1.5/k=0.4.

The fluorophore used is phycoerythrin, absorption=543 nm andemission=580 nm; its altitude with respect to the surface is 10 nm. Thefluorophore is immersed in a liquid environment.

Two different orientations are taken into account for the dipoles: oneparallel and the other perpendicular to the surface. The thickness ofthe polymer is varied and the signal emitted into the medium surroundingthe fluorophore is calculated (case of a very open microscope for theanalysis of fluorescence).

FIG. 17 is a diagram of the dipolar modelling of a fluorophore at thesurface: it represents a dipole with a dipole moment m placed above asurface x. Three media are distinguished: the environment of the dipole,the area 1, in this instance the superstrate; a stack of thin layer(s),the area 2; and the substrate, the area 3. n₁, ε₁; n₂, ε₂ and n₃, ε₃ arerespectively the index and the dielectric permittivity of the areas 1, 2and 3; d represents the distance from the dipole to the surface of thepolymer and t represents the thickness of the polymer.

FIGS. 18 a) and 18 b) represent modellings (signal (ua)=f(thickness ofthe polymer) (in nm)) of the fluorescence signal of a fluorophore dipoleplaced at the surface of an organic film according to two orientations,parallel (FIG. 18 a)) and perpendicular (FIG. 18 b)) to the surface. Inthese figures, case 1: n=1.5 and k=0.02; case 2: n=1.5 and k=0.003; case3: n=1.5 and k=0.0 (extreme value); the case 4 models a typicalconducting organic film, such as, for example, the films obtained fromdiazonium salts (samples 0101Pt6 and 0101Pt14): n=1.5 and k=0.4.

In the light of the results represented in the appended FIGS. 18 a) and18 b), an increase of approximately 25 with regard to the signal isobserved with the present invention using a “MAN” film with a parallelorientation of fluorophores, the most probable case because of thephenomenon of photoselection, and of approximately 100 for aperpendicular orientation.

The predictions of this model are now tested in a simple way byexamining the fluorescence signal of a drop of fluorescent label on afilm of PMAN (AuMAN11 sample) deposited on a gold substrate and bycomparing it with the same signal with regard to a polypyrrole film withthe same thickness deposited on a gold substrate. The fluorophoreemployed is 1 μM Cy₃dctp (trade mark) (Amersham). The volume of thedrops is 0.5 μl.

FIGS. 19 a) and 19 b) are diagrammatic representations of the goldsubstrates coated, on the one hand, with a film of polypyrrole (FIG. 19a)) and, on the other hand, with an insulating organic polymer film(ManAu11) (FIG. 19 b)) in accordance with the present invention.

FIGS. 20 a), b) and c) are negatives (objective ×50; exposure time: 200ms) of the fluorescence on the area of gold (FIG. 20 a)), on the film ofpolypyrrole (FIG. 20 b)) and on the film of polymethacrylonitrile (MAN)which are obtained in this example.

The highly luminous points on the three photographs are eitheragglomerates of fluorophores, or fluorescent dust. These points do notcorrespond to fluorophores close to the surface, that is to say a fewnm, which is the case with biochips, for example, but to balls from afew tens to a few hundreds of nm. It is therefore not with regard tothese light points that the comparison has to be carried out but withregard to the “continuous background” which surrounds these points.

It is observed that, with identical exposure times, photographicreceptors of the same sensitivity, the same magnifications achieved withthe same optics, and even though the concentration of label is very low(1 μm), the fluorophores which are in “direct” contact with the surfaceare not “extinguished” in the case of the AuMAN11 samples (negative c)),whereas they are extinguished over the areas of gold (negative a)) andover the area of the conducting organic coating (negative b)). Thus, inthis instance, the superior properties of transparency of the insulatingfilms of PMAN with regard to the conducting films are directly observed,as illustrated in these FIG. 20.

Example 3 Production of a Deposit of Polymethacrylonitrile Film byElectroinitiation in the Presence of Diazonium Salts

A deposit of polymethacrylonitrile films is produced, on the same goldsurfaces as those of Example No. 1, by electroinitiation starting fromdiazonium salts. These films are obtained by electroreduction of a 2.5Msolution of methacrylonitrile in anhydrous acetonitrile in the presenceof 0,05 M tetraethylammonium perchlorate (TEAP) as supportingelectrolyte and of 10⁻³M 4-nitrophenyldiazonium tetrafluoroborate. Theelectroreduction takes place under voltametric conditions between +0.3and −1.5 V/(Ag⁺/Ag) at 100 mV/s, in nonseparated compartments, with aplatinum counterelectrode with a high surface area. The number of sweepsis adjusted so as to obtain films with a thickness of the order of 100nm.

The optical measurements reveal a coefficient k<0.02, as observed withthe polymethacrylonitrile films obtained by direct electrografting (cf.Table 4).

Example 4 Procedure for Monitoring Reproducibility

Polymethacrylonitrile films of variable thickness peppered withphycoerythrin are manufactured on entirely identical gold strips. Forthis, the gold strips are polarized under voltametric conditions between+0.3 and −1.0 V/(Ag⁺/Ag) at a 100 mV/s, in nonseparated compartments,with a platinum counterelectrode with a high surface area, in a solutioncontaining 2.5M methacrylonitrile in anhydrous acetonitrile, 0,05 Mtetraethylammonium perchlorate (TEAP) as supporting electrolyte, 10⁻³M4-nitrophenyldiazonium tetrafluoroborate and 1 μM phycoerythrin. Thethickness of the films obtained, measured by ellipsometry, is 50±5 nm,i.e. an accuracy of 10%.

Each strip is subsequently irradiated at 543 nm and the resultingintensity of fluorescence is measured at 580 nm. Identical intensitiesof fluorescence are measured on all the strips, with a dispersion of themeasurements of less than 15%.

Example 5 Quality Control Procedure

PMAN (polymethacrylonitrile) films of increasing thickness are depositedaccording to the procedure of Example 4 on 1 cm² gold strips. Thevarious thicknesses, between 5 and 150 nm, are obtained by varying thenumber of voltametric sweeps.

The fluorescence measurement on the coatings obtained makes it possibleto plot a calibration curve giving the thickness as a function of theintensity/cm².

A PMAN deposit with a thickness of 84 nm (measurement by ellipsometry)is subsequently produced on a 5 cm² gold surface. The fluorescencemeasurement indicates, from the calibration curve, a thickness of 75 nm,which makes it possible to validate proper control without directlymeasuring the thickness.

LIST OF REFERENCES

-   [1] Arwin et al., Synthetic Metals, 6, 1983: “Dielectric Function of    Thin Polypyrrole and Prussian Blue Films by Spectroscopic    Ellipsometry”.-   [2] Kim et al., Journal of the Electrochemical Society, 138(11),    1991: “Real Time Spectroscopic Ellipsometry: In Situ    Characterization of Pyrrole Electropolymerization”.-   [3] Kim et al., Bulletin of the Korean Chemical Society, 17(8),    1996: “Polypyrrole Film Studied by Three-Parameter Ellipsometry”.-   [4] Guedon et al., Analytical Chemistry, 22, 6003-6009, 2000:    “Characterization and Optimization of a Real-Time, Parallel,    Label-Free, Polypyrrole-Based DNA Sensor by Surface Plasmon    Resonance Imaging”.

1-16. (canceled)
 17. A substrate coated with a film, wherein the film isan electric insulator organic polymer which is transparent in at leastone wavelength range and wherein the film is combined with a label whichemits at least in the wavelength range.
 18. The substrate of claim 17,wherein the film comprises an insulating polymer selected from vinylpolymers.
 19. The substrate of claim 17, wherein the film comprises aninsulating polymer selected from the group consisting of crosslinked ornoncrosslinked polymers of acrylonitrile, methacrylonitrile, methylmethacrylate, ethyl methacrylate, propyl methacrylate, butylmethacrylate, hydroxyethyl methacrylate, hydroxypropyl methacrylate,cyano acrylate, acrylic acid, methacrylic acid, styrene,para-chlorostyrene, N-vinylpyrrolidone, and vinyl halides.
 20. Thesubstrate of claim 17, wherein the film comprises a crosslinked ornoncrosslinked insulating polymer selected from the group consisting ofpolyacrylamides, polymers of isoprene, of ethylene, of propylene, ofethylene oxide and molecules comprising strained rings, of lactic acidand of its oligomers, of lactones, of ε-caprolactone, of glycolic acidand of aspartic acid, polyamides, polyurethanes, parylene and polymersbased on substituted parylene, oligopeptides and proteins, and theprepolymers, macromers and telechelics based on these polymers, and thecopolymers and/or blends which can be formed from the monomers of thesepolymers and from these polymers themselves.
 21. The substrate of claim17, wherein the film is a vinyl polymer obtained by polymerization of amonomer of formula (I):

in which R₁, R₂, R₃ and R₄ are independently selected from the groupconsisting of hydrogen atoms, alkanes, alkenes, alkynes, amides,aldehydes, ketones, carboxylic acids, esters, acid halides, anhydrides,nitriles, amines, thiols and phosphates, ethers, homo- and heterocyclicaromatics and any cyclic group comprising these functional groups, andany group carrying several of these functional groups; or of a mixtureof different monomers of formula (I).
 22. The substrate of claim 17,wherein the film is polymethacrylonitrile or poly(methyl methacrylate).23. The substrate of claim 17, wherein the label is selected from thegroup consisting of a fluorescent label, a phosphorescent label, and achemiluminescent label. 24-33. (canceled)
 34. A process for thedetection of a chemical entity comprising: providing a substrate coatedwith a film, wherein the film is an electric insulator organic polymerwhich is transparent in at least one wavelength range, wherein the filmis functionalized with recognition molecules for the chemical entity tobe detected; providing a sample to be analyzed, the sample beinglabelled with a label which emits at least in the wavelength range;bringing the sample into contact with the substrate such that thechemical entity to be detected will pair with the recognition moleculesof the film; and detecting for the label.
 35. The process of claim 34,wherein the chemical entity to be detected is DNA.
 36. The process ofclaim 34, wherein the label is selected from the group consisting of afluorescent label, a phosphorescent label, and a chemiluminescent label.37. A biochip comprising: a substrate having deposited thereon a filmcomprising an electrical insulator organic polymer functionalized by abiological probe capable of bringing about pairing with an entityfunctionalized with a fluorescent, phosphorescent or chemiluminescentlabel, wherein the film is transparent in at least one wavelength rangesuitable for detection of the fluorescent, phosphorescent orchemiluminescent label.
 38. The biochip of claim 37, wherein the film isa film of a vinyl polymer.
 39. The biochip of claim 37, wherein the filmis a vinyl polymer obtained by polymerization of a monomer of thefollowing formula (I):

in which R₁, R₂, R₃ and R₄ are independently selected from the groupconsisting of hydrogen atoms, alkanes, alkenes, alkynes, amides,aldehydes, ketones, carboxylic acids, esters, acid halides, anhydrides,nitrites, amines, thiols and phosphates, ethers, homo-and heterocyclicaromatics and any cyclic group comprising these functional groups, andany group carrying several of these functional groups; or of a mixtureof different monomers of formula (I).
 40. The biochip of claim 37,wherein the biological probe is DNA.
 41. The biochip of claim 37,wherein the entity functionalized with a fluorescent, phosphorescent orchemiluninescent label is a chemical, biochemical or biological entity.42. The biochip of claim 41, wherein the entity functionalized with afluorescent, phosphorescent or chemiluminescent label is a chemicalentity.
 43. The biochip of claim 42, wherein the chemical entity is DNA.44. The biochip of claim 41, wherein the entity functionalized with afluorescent, phosphorescent or chemiluminescent label is a biologicalentity.
 45. The biochip of claim 41, wherein the entity functionalizedwith a fluorescent, phosphorescent or chemiluminescent label is anucleic acid, a protein, an antigen, an antibody or a synthetic organicmolecule.
 46. A process for the detection of a sample, comprisingproviding a biochip comprising a substrate having deposited thereon afilm having a thickness of less than one micrometer which is anelectrical insulator organic polymer which is transparent in at leastone wavelength range; providing a sample; labeling the sample with alabel which emits at least in the one said wavelength range; anddetecting the sample with the biochip.
 47. The process of claim 46wherein the film is a film of a vinyl polymer.
 48. The process of claim46 wherein the film is a vinyl polymer obtained by polymerization of amonomer of the following formula (I):

in which R₁, R₂, R₃ and R₄ are independently selected from the groupconsisting of hydrogen atoms, alkanes, alkenes, alkynes, amides,aldehydes, ketones, carboxylic acids, esters, acid halides, anhydrides,nitriles, amines, thiols and phosphates, ethers, homo-and heterocyclicaromatics and any cyclic group comprising these functional groups, andany group carrying several of these functional groups; or of a mixtureof different monomers of formula (I).
 49. The process of claim 46wherein the electrical insulator organic polymer is functionalized by abiological probe capable of bringing about pairing with a chemical,biochemical or biological entity functionalized with a fluorescent,phosphorescent or chemiluminescent label.
 50. The process of claim 49wherein the sample is labeled with a label which is selected from thegroup consisting of a fluorescent label, a phosphorescent label, and achemiluminescent label.
 51. The process of claim 46 wherein thebiological probe is DNA.
 52. A process for the detection of a chemicalentity comprising: providing a film comprising an electrical insulatororganic polymer which is transparent in at least one emission wavelengthrange; providing a sample of a chemical entity to be detected, thesample being labeled with a label which emits in at least the oneemission wavelength range; and detecting for the label of the sample ofthe chemical entity.
 53. The process of claim 52, wherein the filmcomprising an electrical insulator organic polymer is functionalizedwith recognition molecules for the chemical entity.
 54. The process ofclaim 52, wherein the chemical entity is DNA.
 55. The process of claim52, wherein the film is a vinyl polymer obtained by polymerization of amonomer of the following formula (I):

in which R₁, R₂, R₃ and R₄ are independently selected from the groupconsisting of hydrogen atoms, alkanes, alkenes, alkynes, amides,aldehydes, ketones, carboxylic acids, esters, acid halides, anhydrides,nitrites, amines, thiols and phosphates, ethers, homo-and heterocyclicaromatics and any cyclic group comprising these functional groups, andany group carrying several of these functional groups; or of a mixtureof different monomers of formula (I).
 56. The process of claim 52,wherein the film is of polymethacrylonitrile or ofpoly(methylmethacrylate).
 57. The process of claim 52, wherein the labelis selected from the group consisting of a fluorescent label, aphosphorescent label and a chemiluminescent label.
 58. A process for thedetection of a chemical entity comprising: providing a means for thedetection of the chemical entity comprising a film comprising anelectrical insulator organic polymer which is transparent in at leastone emission wavelength range; providing a sample of a chemical entityto be detected, the sample being functionalized with a label which emitsin at least the one emission wavelength range; and detecting for thelabel of the sample of the chemical entity.
 59. The process of claim 58,wherein the detection means is a biochip.
 60. The process of claim 58,wherein the film is a vinyl polymer obtained by polymerization of amonomer of the following formula (I):

in which R₁, R₂, R₃ and R₄ are independently selected from the groupconsisting of hydrogen atoms, alkanes, alkenes, alkynes, amides,aldehydes, ketones, carboxylic acids, esters, acid halides, anhydrides,nitrites, amines, thiols and phosphates, ethers, homo-and heterocyclicaromatics and any cyclic group comprising these functional groups, andany group carrying several of these functional groups; or of a mixtureof different monomers of formula (I).
 61. The process of claim 58,wherein the film is of polymethacrylonitrile or ofpoly(methylmethacrylate).
 62. The process of claim 58, wherein the labelis selected from the group consisting of a fluorescent label, aphosphorescent label and a chemiluminescent label.
 63. A process forquality control, certification or authentication of an objectcomprising: obtaining the object by combining a film comprising anelectrical insulator organic polymer which is transparent in at leastone emission wavelength range with a label which emits in at least theone emission wavelength range; and measuring the intensity of theemission of the at least one wavelength range of the object.
 64. Theprocess of claim 63, wherein the film is a vinyl polymer obtained bypolymerization of a monomer of the following formula (I):

in which R₁, R₂, R₃ and R₄ are independently selected from the groupconsisting of hydrogen atoms, alkanes, alkenes, alkynes, amides,aldehydes, ketones, carboxylic acids, esters, acid halides, anhydrides,nitriles, amines, thiols and phosphates, ethers, homo-and heterocyclicaromatics and any cyclic group comprising these functional groups, andany group carrying several of these functional groups; or of a mixtureof different monomers of formula (I).
 65. The process of claim 63,wherein the film is of polymethacrylonitrile or ofpoly(methylmethacrylate).
 66. The process of claim 63, wherein the labelis selected from the group consisting of a fluorescent label, aphosphorescent label and a chemiluminescent label.
 67. The process ofclaim 66, comprising measuring the intensity of fluorescence emittedover the emission wavelength of the fluorescent label.